It’s quite necessary to fabricate reproducible and uniform surface-enhanced Raman scattering (SERS) composite substrate with high enhancement factor simply. Therefore, in this work, in order to obtain the SERS substrates with optimized size and gap, Ag films with different thickness deposited by magnetron sputtering and following annealing are performed. The results both elucidate the function relationship between the size, gap of nanoparticles and the thickness of Ag films, and ascertain the optimized parameters for silver nanoparticles on the basis of finite-difference time-domain simulation, the SERS signal using graphene and Rhodamine 6G (R6G) as probe molecules for Ag-NPs/Si substrates. Moreover, our findings highlight the Ag NPs with optimized size and gap as SERS substrates present high reproducibility and uniformity.